LOCUS       sequence-425902-        5784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human EIF3B fragment V186 to T685. 
            N-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5784)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova YA, Baghaie AJ, Beldar S, Bolotokova A, Centrella
            PA, Chau I, Clark MA, Cuozzo JW, Dehghani-Tafti S, Disch JS, Dong A,
            Dumas A, Feng JA, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green SR,
            Guie MA, Guilinger JP, Harms N, Herasymenko O, Houliston S, 
            Hutchinson A, Kearnes S, Keefe AD, Kimani SW, Kramer T, Kutera M, 
            Kwak HA, Lento C, Li Y, Liu J, Loup J, Machado RAC, Mulhern CJ, 
            Perveen S, Righetto GL, Riley P, Shrestha S, Sigel EA, Silva M, 
            Sintchak MD, Slakman BL, Taylor RD, Thompson J, Torng W, 
            Underkoffler C, von Rechenberg M, Walsh RT, Watson I, Wilson DJ, 
            Wolf E, Yadav M, Yazdi AK, Zhang J, Zhang Y, Santhakumar V, Edwards 
            AM, Barsyte-Lovejoy D, Schapira M, Brown PJ, Halabelian L, 
            Arrowsmith CH
  TITLE     A Target Class Ligandability Evaluation of WD40 Repeat-Containing 
            Proteins.
  JOURNAL   J Med Chem. 2025 Jan 23;68(2):1092-1112. doi: 
            10.1021/acs.jmedchem.4c02010. Epub 2024 Nov 4.
  PUBMED    39495097
REFERENCE   2  (bases 1 to 5784)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acs.jmedchem.4c02010"; journalName: "J Med Chem"; date: 
            "2025-01-23- 23"; volume: "68"; issue: "2"; pages: "1092-1112"
COMMENT     Sequence Label: pBiT3.1-N_EIF3B:186-685
FEATURES             Location/Qualifiers
     source          1..5784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2847..2968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2858..2877)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3083..3116)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3126..3173
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3205..3999
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3259..3278)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3869..3888
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4157..4745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4237..4256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4871..4919
                     /note="synthetic polyadenylation signal"
     CDS             complement(4943..5341)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5338..5347
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5372..5729)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5386..5521
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5538..5729
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5686..5706
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaatctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact ataggggtta gcgatcgcca tggtgagcgg
     1201 ctggcggctg ttcaagaaga ttagcgggag ctccggtggc tcgagcggtg tgattgtagt
     1261 ggacaatgtc cctcaggtgg gacccgaccg acttgagaaa ctcaaaaatg tcatccacaa
     1321 gatcttttcc aagtttggga aaatcacaaa tgatttttat cctgaagagg atgggaagac
     1381 aaaagggtat attttcctgg agtacgcgtc ccctgcccac gctgtggatg ctgtgaagaa
     1441 cgccgacggc tacaagcttg acaagcagca cacattccgg gtcaacctct ttacggattt
     1501 tgacaagtat atgacgatca gtgacgagtg ggatattcca gagaaacagc ctttcaaaga
     1561 cctggggaac ttacgttact ggcttgaaga ggcagaatgc agagatcagt acagtgtgat
     1621 ttttgagagt ggagaccgca cttccatatt ctggaatgac gtaaaagacc ctgtctcaat
     1681 tgaagaaaga gcgagatgga cagagacgta tgtgcgttgg tctcctaagg gcacctacct
     1741 ggctaccttt catcaaagag gcattgctct atggggggga gagaaattca agcaaattca
     1801 gagattcagc caccaagggg ttcagcttat tgacttctca ccttgtgaaa ggtacctggt
     1861 gacctttagc cccctgatgg acacgcagga tgaccctcag gccataatca tctgggacat
     1921 ccttacgggg cacaagaaga ggggttttca ctgtgagagc tcagcccatt ggcctatttt
     1981 taagtggagc catgatggca aattctttgc cagaatgacc ctggatacgc ttagcatcta
     2041 tgaaactcct tctatgggtc ttttggacaa gaagagtttg aagatctctg ggataaaaga
     2101 cttttcttgg tctcctggtg gtaacataat cgccttctgg gtgcctgaag acaaagatat
     2161 tccagccagg gtaaccctga tgcagctccc taccaggcaa gagatccgag tgaggaacct
     2221 gttcaatgtg gtggactgca agctccattg gcagaagaac ggagactact tgtgtgtgaa
     2281 agtagatagg actccgaaag gcacccaggg tgttgtcaca aattttgaaa ttttccgaat
     2341 gagggagaaa caggtacctg tggatgtggt cgagatgaaa gaaaccatca tagcctttgc
     2401 ctgggaacca aatggaagta agtttgctgt gctgcacgga gaggctccgc ggatatctgt
     2461 gtctttctac cacgtcaaaa acaacgggaa gattgaactc atcaagatgt tcgacaagca
     2521 gcaggcgaac accatcttct ggagccccca aggacagttc gtggtgttgg cgggcctgag
     2581 gagtatgaac ggtgccttag cgtttgtgga cacttcggac tgcacggtca tgaacatcgc
     2641 agagcactac atggcttccg acgtcgaatg ggatcctact gggcgctacg tcgtcacctc
     2701 tgtgtcctgg tggagccata aggtggacaa cgcgtactgg ctgtggactt aatctagagg
     2761 tcaagcttcg ggggtttaaa cctgcaggca tgcaagctga tttggctgct gccaccgctg
     2821 agcaaggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     2881 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2941 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3001 ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa
     3061 aatcttaatt aagacttcta atataacttc gtatagcata cattatacga agttatggtt
     3121 ccactgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     3181 ggaactaaac ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg
     3241 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3301 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3361 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3421 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3481 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3541 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3601 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3661 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3721 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3781 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3841 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3901 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3961 cgcatcgcct tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg
     4021 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4081 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4141 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4201 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4261 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4321 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4381 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4441 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4501 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4561 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4621 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4681 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4741 ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4801 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt
     4861 ccggaatcgt cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg
     4921 cggccatcgt gatggctaga gtttagccct cccacacata accagagggc agcaattcac
     4981 gaatcccaac tgccgtcggc tgtccatcac tgtccttcac tatggctttg atcccaggat
     5041 gcagatcgag aagcacctgt cggcaccgtc cgcaggggct caagatgccc ctgttctcat
     5101 ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc agcagcagca gtgcccagca
     5161 ccacgagttc tgcacaaggt cccccagtaa aatgatatac attgacacca gtgaagatgc
     5221 ggccgtcgct agagagagct gcgctggcga cgctgtagtc ttcagagatg gggatgctgt
     5281 tgattgtagc cgttgctctt tcaatgaggg tggattcttc ttgagacaaa ggcttggcca
     5341 tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc
     5401 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     5461 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     5521 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     5581 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     5641 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     5701 cattccacag ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg
     5761 ttatttcagg ccatggtgct gcgc
//
